Transendothelial transport mediated modulations to the High Density Lipoprotein

跨内皮转运介导的高密度脂蛋白调节

基本信息

批准号：
10245152
负责人：
VASANTHY NARAYANASWAMI
金额：
$ 11.06万
依托单位：
CALIFORNIA STATE UNIVERSITY LONG BEACH
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10245152
关键词：
ATP-Binding Cassette Transporters Address Amphipathic Alpha Helix Antiatherogenic Antioxidants Apolipoprotein A-I Apolipoproteins Arterial Fatty Streak Arterial Intimas Arylesterase Attention Biomedical Research Blood Vessels Cardiovascular Diseases Cardiovascular system Cellular biology Cholesterol Collaborations Communities Development Endothelial Cells Evaluation Exhibits Fluorescent Probes Funding Future Goals High Density Lipoprotein Cholesterol High Density Lipoproteins Impairment In Vitro Joints Knowledge Leadership Life Style Light Lipids Lipoproteins Literature Location Low Density Lipoprotein oxidation Mass Spectrum Analysis Mediating Mentors Mentorship Metabolism Modification Molecular Conformation Outcome Particle Size Patients Pharmacology Plasma Post-Translational Protein Processing Property Proteins Proteomics Publications Publishing Reporting Research Research Training Risk Role Structure Structure-Activity Relationship Students Testing Ursidae Family antioxidant enzyme apolipoprotein E-3 atheroprotective base biomarker identification cardiovascular disorder risk cardiovascular risk factor conformational alteration flexibility gene therapy improved insight lipidomics macrophage monolayer particle prevent reconstitution response skills symposium training opportunity transcytosis

项目摘要

Title: Transendothelial transport mediated modulations to the High Density Lipoprotein Summary Despite the vast amount of literature that indicates low plasma levels of HDL-cholesterol (HDL-C) is a risk factor for cardiovascular disease (CVD), the last decade has seen a paradigm shift in the concept that it may not be HDL-C levels per se but the functionality of HDL that is a determining factor in CVD. The shift in focus is in light of several studies that show that neither pharmacological nor genetic intervention to increase HDL-C levels, lower the risk for CVD. Thus, there is a pressing need to fill the gap in knowledge regarding the role of HDL in CVD from a mechanistic perspective and understand structure-function relationships in HDL. In the current proposal, we hypothesize that apolipoprotein (apo) AI and apoE3, two critical apos on HDL, undergo structural alterations and post translational modifications (PTM) and that HDL undergoes particle modulation as a consequence of transendothelial transport from plasma to the arterial intima with significant functional penalties. We will test this hypothesis by interrogating structure-function changes to the HDL with two specific aims: (1) Identify structural and proteomic alterations to the HDL associated with transendothelial transport across aortic endothelial cells, and, (2) Determine changes in HDL function following transendothelial transport. We will carry out spectroscopic analysis of reconstituted HDL (rHDL) bearing spatially sensitive fluorescent probes at flexible locations on apoAI or apoE3 to obtain insight into conformational alterations following transcytosis. Based on previous findings about oxidatively modified apoAI in atherosclerotic plaques, we postulate that the apos are susceptible to oxidative modification during transcytosis. To address this, we will determine PTM in transcytosed HDL, specifically on apoAI, apoE3 and selected proteins involved in lipoprotein metabolism by mass spectrometry. We will also perform lipoproteomic analysis to identify changes to the protein and lipid composition of transcytosed HDL. In an independent but complementary approach, we will examine changes in two major functional effects of transcytosed HDL: its ability to promote cholesterol efflux from macrophages, and, its antioxidant activity. Completion of these studies will significantly advance our understanding of the relationship between structure/composition and the atheroprotective effect of HDL at the vascular wall. The expected outcome of the proposed studies is to have a deeper understanding of modulations in HDL that render it dysfunctional. Establishing this relationship will aid in development of HDL- based therapies and identification of biomarkers of CVD risk. The research enhancement objectives of the PI are to: (i) develop expertise in HDL and endothelial cell biology, (ii) increase competitiveness to apply for major external funding by publishing research findings, (iii) strengthen existing and develop new collaborations, submit joint publications and lay the groundwork for developing proposals, and, (iv) establish mentoring, networking, and leadership skills, and offer research training opportunities and mentorship for students from diverse backgrounds in biomedical research.

标题：对高密度脂蛋白的跨内皮传输介导的调节概括尽管有大量文献表明血浆水平较低的HDL-胆固醇（HDL-C）是一种风险心血管疾病（CVD）的因素，在过去的十年中，概念可能发生了范式转变不是HDL-C水平本身，而是HDL的功能，这是CVD中的决定因素。重点的转变是鉴于几项研究表明，既不增加HDL-C的药理和遗传干预措施水平，降低CVD的风险。因此，迫切需要填补有关的知识的差距从机械角度来看，CVD中的HDL并了解HDL中的结构 - 功能关系。在当前的建议中，我们假设载脂蛋白（APO）AI和APOE3，HDL上的两个关键APO，经历结构改变和翻译后修改（PTM），HDL经历粒子从血浆到动脉内膜的跨内皮转运导致的调节功能罚款。我们将通过询问与HDL的结构 - 功能变化来检验该假设两个具体的目的：（1）确定与跨内皮相关的HDL的结构和蛋白质组学改变跨主动脉内皮细胞的运输，（2）确定跨内皮后HDL功能的变化运输。我们将对具有空间敏感的重构HDL（RHDL）进行光谱分析 APOAI或APOE3上柔性位置的荧光探针，以了解构象改变跨胞菌病后。基于先前关于动脉粥样硬化斑块中氧化修饰的apoAI的发现，我们假设APO易于转胞胞菌病期间氧化修饰。为了解决这个问题，我们将确定跨性别HDL中的PTM，特别是在ApoAI，APOE3和参与的选定蛋白质谱法代谢脂蛋白代谢。我们还将执行脂肪蛋白质组学分析以识别变化转化HDL的蛋白质和脂质组成。在一种独立但互补的方法中，我们将检查跨性别HDL的两个主要功能效应的变化：促进胆固醇的能力巨噬细胞的外排及其抗氧化活性。这些研究的完成将大大推动我们的了解HDL在结构/组成与HDL的动脉保护作用之间的关系血管壁。拟议研究的预期结果是对在HDL中调制它，使其功能失调。建立这种关系将有助于发展HDL- 基于CVD风险的生物标志物的疗法和鉴定。 PI的研究增强目标是：（i）在HDL和内皮细胞中发展专业知识生物学，（ii）通过发布研究结果提高竞争力，以申请主要的外部资金，（iii）加强现有并开发新的合作，提交联合出版物并为制定建议，以及（iv）建立指导，网络和领导能力，并提供研究来自生物医学研究背景的学生的培训机会和指导。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Apolipoprotein E LDL receptor-binding domain-containing high-density lipoprotein: a nanovehicle to transport curcumin, an antioxidant and anti-amyloid bioflavonoid.

DOI：
10.1016/j.bbamem.2010.09.007
发表时间：
2011-01
期刊：
Biochimica et biophysica acta
影响因子：
0
作者：
Khumsupan P;Ramirez R;Khumsupan D;Narayanaswami V
通讯作者：
Narayanaswami V

Targeted intracellular delivery of resveratrol to glioblastoma cells using apolipoprotein E-containing reconstituted HDL as a nanovehicle.

DOI：
10.1371/journal.pone.0135130
发表时间：
2015
期刊：
PloS one
影响因子：
3.7
作者：
Kim SH;Adhikari BB;Cruz S;Schramm MP;Vinson JA;Narayanaswami V
通讯作者：
Narayanaswami V

Ordered opening of LDL receptor binding domain of human apolipoprotein E3 revealed by hydrogen/deuterium exchange mass spectrometry and fluorescence spectroscopy.

氢/氘交换质谱和荧光光谱显示人载脂蛋白 E3 的 LDL 受体结合域有序开放。

DOI：
10.1016/j.bbapap.2018.08.005
发表时间：
2018
期刊：
Biochimica et biophysica acta. Proteins and proteomics
影响因子：
0
作者：
Yang,Liping;Hernandez,RoyV;Tran,TuyenN;Nirudodhi,Sasidhar;Beck,WendyHJ;Maier,ClaudiaS;Narayanaswami,Vasanthy
通讯作者：
Narayanaswami,Vasanthy

Biochemical and biophysical characterization of recombinant rat apolipoprotein E: similarities to human apolipoprotein E3.

重组大鼠载脂蛋白 E 的生化和生物物理特征：与人载脂蛋白 E3 的相似性。

DOI：
10.1016/j.abb.2012.10.007
发表时间：
2013
期刊：
Archives of biochemistry and biophysics
影响因子：
3.9
作者：
Tran,TuyenN;Kim,SeaH;Gallo,Carlos;Amaya,Max;Kyees,Jessica;Narayanaswami,Vasanthy
通讯作者：
Narayanaswami,Vasanthy

Apolipoprotein E3-mediated cellular uptake of reconstituted high-density lipoprotein bearing core 3, 10, or 17 nm hydrophobic gold nanoparticles.